# US 11,028,149 B2 · Recombinant polypeptide production method

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,028,149 B2
- Document ID: US11028149B2
- Status: Granted
- Kind: B2
- Application number: 14/008,791
- Filed: 2012-03-30
- Published: 2014-01-30
- Granted: 2021-06-08
- Earliest claimed: 2011-04-01
- Assignees: Chugai Seiyaku Kabushiki Kaisha
- Applicants: Hisahiro Tabuchi; Tomoya Sugiyama
- Inventors: Hisahiro Tabuchi; Tomoya Sugiyama
- Web page: https://patentgrep.com/patent/US11028149B2

## CPC

- C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- C07K 16/2866: {against receptors for cytokines, lymphokines, interferons}
- C07K 16/303: {Liver or Pancreas}
- C07K 2317/14: Specific host cells or culture conditions, e.g. components, pH or temperature
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2510/02: Cells for production
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

Provided is a method capable of producing a protein at a high level using a cultured animal cell, comprising culturing a cell that expresses APES (Antibody Production Enhancing Sequence) and into which a DNA encoding a desired polypeptide has been introduced, thereby producing the desired polypeptide. APES contains a nucleotide sequence related to nuclear factor κB inhibitor α (NfkBia) and has a function of decreasing the intracellular expression of NfkBia.

## Claims

### Claim 1

A method of producing an antibody comprising culturing a cell into which a DNA encoding a desired antibody and APES (Antibody Production Enhancing Sequence) have been exogenously introduced by transfection, thereby producing the desired antibody, wherein
the APES is a nucleic acid molecule that increases the ability of the cultured cells to produce the antibody, wherein the APES is selected from nucleic acid molecules each consisting of any of the following nucleotide sequences:
(a) a DNA consisting of the nucleotide sequence of any of SEQ ID NOs:1 to 16 and 29;
(b) a DNA consisting of 19 to 25 nucleotides in the sequence of SEQ ID NO: 2;
(c) an RNA that is a transcript of (a), or (b); and
(d) a DNA or RNA consisting of a sequence that can bind 19 to 25 nucleotides in the sequence of SEQ ID NO: 2 by base pairing.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the APES has been introduced into the cell by an expression vector.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the cell into which the APES has been artificially introduced is a cell in which the transcription of endogenous APES has been activated.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the method further comprises introducing a DNA encoding a taurine transporter.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the method further comprises introducing a DNA encoding cysteine sulfinic acid.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the method further comprises introducing a DNA encoding alanine transferase.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the cell is a Chinese hamster ovary cell.

### Claim 8 (depends on 1)

A method of producing a pharmaceutical composition comprising an antibody, wherein the method comprises producing an antibody by the method according to claim 1, and mixing the antibody with a pharmaceutically acceptable carrier or additive.